Abe Sadanobu, Yasumura Ayako, Tanaka Teruo
Institute of Oceanic Research and Development, Tokai University, Orido 3-20-1, Shimizu-ku, Shizuoka 424-8610, Japan.
J Bacteriol. 2009 May;191(9):3050-8. doi: 10.1128/JB.00049-09. Epub 2009 Feb 27.
Expression of the gene for the extracellular alkaline protease (aprE) of Bacillus subtilis is subject to regulation by many positive and negative regulators. We have found that aprE expression was increased by disruption of the glutamine synthetase gene glnA. The increase in aprE expression was attributed to a decreased in expression of scoC, which encodes a negative regulator of aprE expression. The glnA effect on scoC expression was abolished by further disruption of tnrA, indicating that aprE expression is under global regulation through TnrA. In the scoC background, however, aprE expression was decreased by glnA deletion, and it was shown that the decrease was due to a defect in positive regulation by DegU. Among the genes that affect aprE expression through DegU, the expression of degR, encoding a protein that stabilizes phosphorylated DegU, was inhibited by glnA deletion. It was further shown that the decrease in degR expression by glnA deletion was caused by inhibition of the expression of sigD, encoding the sigma(D) factor, which is required for degR expression. In accordance with these findings, the expression levels of aprE-lacZ in glnA scoC degR and scoC degR strains were identical. These results led us to conclude that glnA deletion brings about two effects on aprE expression, i.e., a positive effect through inhibition of scoC expression and a negative effect through inhibition of degR expression, with the former predominating over the latter.
枯草芽孢杆菌细胞外碱性蛋白酶(aprE)基因的表达受到多种正调控因子和负调控因子的调节。我们发现,谷氨酰胺合成酶基因glnA的缺失会增加aprE的表达。aprE表达的增加归因于scoC表达的降低,scoC编码aprE表达的负调控因子。通过进一步破坏tnrA,消除了glnA对scoC表达的影响,这表明aprE的表达受TnrA的全局调控。然而,在scoC背景下,glnA缺失会降低aprE的表达,并且表明这种降低是由于DegU正调控缺陷所致。在通过DegU影响aprE表达的基因中,编码稳定磷酸化DegU的蛋白质的degR的表达受到glnA缺失的抑制。进一步表明,glnA缺失导致degR表达降低是由于编码σ(D)因子的sigD的表达受到抑制,而sigD是degR表达所必需的。根据这些发现,glnA scoC degR和scoC degR菌株中aprE-lacZ的表达水平是相同的。这些结果使我们得出结论,glnA缺失对aprE表达产生两种影响,即通过抑制scoC表达产生的正效应和通过抑制degR表达产生的负效应,前者比后者占主导地位。